The studies on superinfection of Raji cells with EBV will be performed further. Virus DNA obtained from superinfection will be cleaved by restriction endonucleases, Eco R 1, Hind lll, and Xba and the fragments obtained will be placed on a physical map. The overlapping mapping method by reciprocal digestion of virus DNA with two enzymes will be used. Intact single stranded DNA will be isolated from glycerol gradients and self annealed so as to observe inverted repetitious sequences under an electronmicroscope. Virus mRNA induced after superinfection of Raji cells will be pulse labeled and characterized for the size and location of transcribed sequences on the physical map. Early and late mRNA will be defined by using phosphonoacetic acid which inhibits EBV DNA replication and virus specific RNA in transformed cells will be compared to these two types of mRNA. Proteins synthesized after infection of BJA (EBV genome negative B) cells, as well as superinfected Raji cells with EBV from both HR-1 and B95 cultures, will be analyzed to determine how far the infection proceeds in each system and what causes an abortive infection. DNA binding proteins found in Raji and superinfected Raji cells but not in Simpson (EBV genome free) cells will be further characterized. The possibility of a few genomes being integrated into Raji cell DNA and the integration of one genome in HR-1 #9 cells (one genome/cell) will be studied by a blotting technique which involves gel electrophoresis after restriction enzyme digestion, the transfer of DNA fragments to nitrocellulose filters and hybridization of fragments with virus specific radioactive DNA or cRNA. BIBLIOGRAPHIC REFERENCES: Deinhardt, F., Falk, L., Wolfe, L. G., Schudel, A., Nonoyama, M., Lai, P., Lapin, B., and Yakovleva, L. Susceptibility of Marmosets to Epstein-Barr virus-like Baboon Herpesviruses, Oak Ridge Associated Universities Conference on Marmosets in Experimental Medicine. 1977, in press. Tanaka, A. Suitability of Epstein-Barr Virud DNA Obtained from Superinfected Raji Cells for Complementary RNA Hybridization Studies. J. Gen Virol., 34, 375-379, 1977.